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We sought to develop a co-culture biofilm model to characterize interactions between these two opportunistic pathogens. Upon growth in biofilm-promoting media containing added glucose, fructose, or trehalose, VRE produces sufficient acid to lower the pH and inhibit growth of\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      . We found this effect depended on the carbon source, and that acidification by VRE was necessary and sufficient to suppress\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      growth in liquid medium and in cecal content extracts from germ-free mice. VRE frequently dominates the intestine of patients administered antibiotics, which can predispose to the development of\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      infection. We reasoned that it may be possible to suppress\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      growth during co-infection with VRE by supplementing the mouse diet with a fermentable sugar. A VRE-dominated gut microbiota may convert the sugar to acid, lower the pH, and reestablish colonization resistance to\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      . Supplementation of the diet of VRE-colonized mice with high levels of fructose neither resulted in a lower pH nor did it prevent colonization by\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      . Taken together, these data suggest that VRE can suppress growth of\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      by organic acid production in a carbon source-dependent manner\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      ; however, the mammalian intestine may require sophisticated approaches to lower pH therapeutically.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/iai.00159-26","type":"journal-article","created":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T13:00:47Z","timestamp":1778677247000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Acidification-dependent suppression of\n                    <i>C. difficile<\/i>\n                    by pathogenic and commensal 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